Northern Illinois University, Dekalb, IL 60115
We are interested in quantitatively measuring the development of autophagosomes in irradiated human brain cancer, glioblastoma multiforme U251, cells after exposure to g radiation. Our experiments involved measuring the intensity or “brightness” emitted by fluorescently stained cells, which is directly related to the number and size of autophagosomes in the cells. We grew U251 GBM cells in tissue culture and irradiated the cells using g radiation from a cesium source. Cells were then stained with acridine orange fluorescent dye, and the “brightness” emitted by individual cells was measured 3 and 7 days after irradiation. Those cells subjected to g rays displayed a greater “brightness” after AO staining compared to unirradiated samples. A clear increase in cell “brightness” from the 3 day to the 7 day time period was also observed. With this method, we are able to determine which irradiation therapies will cause the greatest extent of autophagy in each cell line. Ultimately, we are working on developing irradiation therapies that will prove more effective against cancerous human GBM.
[Abstract (DOCX)]